Preclinical toxicological evaluation of Aloe vera health drinks in wistar rats

Human consumption of Aloe vera as a beverage has recently increased in popularity. These benefits are controversial with some sources pointing out that the putative effects of aloe are unsupported by clinical studies; it is important that marketed products be tested for toxicities following oral consumption. Hence this study was designed to evaluate the toxicological effect of marketed aloe health drinks. Thirty either sex Wistar rats (200-300gm) were enrolled in this study and are divided into 5 groups. Group I receives Normal saline serves as vehicle control, Group II and III receives Product A- Low dose (0.5 ml twice daily, p.o) and High dose (1.0 ml twice daily, p.o) respectively. Group IV and V receives Product B- Low dose (0.5 ml twice daily, p.o) and High dose (1.0 ml twice daily, p.o) respectively. Weekly body weight and daily feed intake were measured. On 28th day total urine output volume, faecal consistency, Haematological, biochemical, and organ weight were measured to assess the toxicity of aloe health drinks. The result of this study shows that continuous usage of aloe health drinks showed milder weight reduction, significant improvement in erythropoiesis also it increases the WBC count and increases the weight of spleen it may confirm the immune modulatory effect of aloe health drink. At the higher doses, it increased the SGOT, SGPT, serum urea and creatinine it may lead to the hepatotoxicity and nephrotoxicity. In gastrointestinal tract on prolonged uses, it produced few lesions and diarrhoea. It might be concluded that prolonged consumption of unprocessed aloe health drink contains latex, an ingredient which has many health risks associated with it. So it can aggravate health problems

Dual Fungal Infection of Aspergillosis and Mucormycosis in a COVID-19 Patient: A Rare Case Report

Coronavirus disease 2019 (COVID-19) infections can be related to vast spectrum of co-existent bacterial and fungal infections. A 49-year-old diabetic male was admitted with a history of fever, cough and breathlessness since 5 days. He developed persistent headache with right sided purulent nasal discharge. Relevant histo-pathological, biochemical, microbiological and imaging studies were performed which proved it to be a dual infection of Aspergillosis and Mucormycosis. We present one such case in a COVID-19 patient to highlight its unusual clinical features along with the diagnostic and therapeutic challenges

Isolated Intracranial Myeloid Sarcoma Occurring as Relapse in Acute Myeloid Leukemia

Myeloid sarcoma (MS) or chloroma is a rare extramedullary tumor composed of extramedullary proliferation of blasts of granulocytic, monocytic, erythroid, or megakaryocytic lineage occurring at sites outside the bone marrow. MS occurs in 2%–8% of patients with acute myeloid leukemia (AML), sometimes it occurs as the presenting manifestation of relapse in a patient in remission. We describe the case of a young male with AML in remission for 6 years presenting with central nervous system symptoms. Magnetic resonance imaging showed an extra-axial altered intensity lesion in the parasagittal parietal region, infiltrating anterosuperiorly into anterior falx, and posterosuperior aspect of the superior sagittal sinus. A biopsy from the lesion was diagnostic of MS which was positive for myeloperoxidase. He did not have any other sites of disease. He has received chemotherapy with FLAG (Fludarabine, Cytosine arabinoside) followed by cranial irradiation and is in complete remission

Mucosal Tolerance to a Combination of ApoB and HSP60 Peptides Controls Plaque Progression and Stabilizes Vulnerable Plaque in Apob^tm2SgyLdlr^tm1Her/J Mice

<div><p>Oral tolerance to auto antigens reduces the development of atherosclerosis in mouse models. However, the effect of immune tolerance to multiple self antigenic peptides in plaque progression and stabilization is not known. We studied the protective effect of mucosal tolerance to peptides from apolipoprotein B (ApoB; 661–680) and heat shock protein 60 (HSP60; 153–163), in combination with diet, in the prevention of atherosclerotic lesion progression and plaque stabilization in ApoB^tm25gyLDLr^tm1Her mice. We found that oral administration of five doses of a combination of ApoB and HSP60 peptides (20 µg/mice/dose) induced tolerance to both the peptides and reduced early plaque development by 39.9% better than the individual peptides (ApoB = 28.7%;HSP60 = 26.8%)(P<0.001). Oral tolerance to combination of peptides along with diet modification arrested plaque progression by 37.6% which was associated with increases in T-regulatory cell and transforming growth factor-β expression in the plaque and peripheral circulation. Reduced macrophage infiltration and tumor necrosis factor-α expression in the plaque was also observed. Tolerance with continued hypercholesterolemia resulted in 60.8% reduction in necrotic core area suggesting plaque stabilization, which was supported by reduction in apoptosis and increased efferocytosis demonstrated by greater expression of receptor tyrosine kinase Mer (MerTK) in the plaque. Tolerance to the two peptides also reduced the expression of matrix metalloproteinase 9, tissue factor, calprotectin, and increased its collagen content. Our study suggests that oral tolerance to ApoB and HSP60 peptide combination induces CD4⁺ CTLA4⁺ Tregs and CD4⁺CD25⁺Foxp3⁺ Tregs secreting TGF-β, which inhibit pathogenic T cell response to both peptides thus reducing the development and progression of atherosclerosis and provides evidence for plaque stabilization in ApoB^tm25gyLDLr^tm1Her mice.</p> </div

Oral Administration of Combination of ApoB and HSP60 Peptides Provides Improved Efficacy against Atherosclerosis Compared to Individual Peptides.

<p>A. Experimental design. B. Representative photomicrographs of EVG stained plaque area and its quantitative analysis in aortic sinus of 18 week old ApoB^tm25gyLDLr^tm1Her mice (n = 10 per group). Scale bar represents 200 µm. C. Percentage of CD25⁺Foxp3⁺ cells (*P<0.05) within the CD4 population in spleen at the end of the study using flow cytometry analysis (n = 6 per group).</p

Oral Tolerance with Continued Hypercholesterolemia Stabilizes Vulnerable Plaque.

<p>A. Experimental design. B. Representative photomicrographs of plaque area stained with EVG and its quantitative analysis in aortic sinus of 26 week old ApoB^tm25gyLDLr^tm1Her mice (n = 10 per group). Scale bar represents 200 µm C. Percentage of acellular necrotic core area in total plaque area. *P = 0.012 D. Representative photomicrographs showing double immunofluorescence staining of aortic sinus sections with CD4 (green) and Foxp3 (red). Scale bar represents 50 µm. Enlarged region to show double immune staining. Scale bar represents 6.25 µm. Right panel: Number of CD4-positive cells/mm² and CD4+ Foxp3+ cells/mm² (n = 9per group). ***P<0.001. E. Percentage of CD25⁺Foxp3⁺ cells (*P<0.009) and CTLA-4 (NS) within the CD4 population in spleen (n = 6 per group). F. Expression of mRNA of Foxp3 (***P = 0.003), CTLA-4 (P = NS), and TGF-β (**P = 0.005) in the ascending aorta quantified by RT-PCR and normalized to GAPDH. Fold-changes in their expression in ApoB+HSP60-tolerized mice relative to controls (n = 4 per group).</p

Model Depicting Oral Tolerance-Induced Prevention and Stabilization of Atherosclerotic Lesion Development.

<p>Oral administration of peptides (ApoB+HSP60) induces a regulatory T-cell response, which is maintained for 10 weeks and results in prevention of early atherosclerotic lesions. Oral administration of peptides in mice with established lesions in combination with diet modification also induces a Treg-cell response with an increase in CD4+CD25+Foxp3 T cells, CTLA-4 expression, and increased TGF-secretion in the lesion and the peripheral circulation. It is likely that both CTLA-4- and TGF-mediated suppression are required for early lesion reduction and inhibition of plaque progression. With continuous hypercholesterolemia, the CTLA-4 expression remains unaltered, while there is an increase in Foxp3 and TGF expression in tolerized mice. We propose that the TGF-mediated decrease in inflammatory activity results in plaque stabilization.</p

Reduction in Plaque Vulnerability Markers in Tolerized Mice.

<p>A. Representative photomicrographs showing immunofluorescence staining of aortic sinus sections with MMP9 (red) and its quantitative analysis (n = 10 per group). *P = 0.035 B. Representative photomicrographs showing immunofluorescence staining of aortic sinus sections with tissue factor (red) and its quantitative analysis (n = 6 per group). *P = 0.028 C. Representative photomicrographs showing immunofluorescence staining of aortic sinus sections with calprotectin (MRP8/14) (red) and its quantitative analysis (n = 6 per group). *P = 0.045 D. Representative photomicrographs of aortic sinus sections stained with alizarin-red S. (n = 6 per group) Arrows indicate calcium depositions and its quantitative analysis. *P = 0.018. Scale bar represents 150 µm. E. Representative photomicrographs of aortic sinus sections stained with Masson’s trichrome and its quantitative analysis (n = 8 per group). **P = 0.002. Lower photomicrograph shows picrosirius staining for collagen content Scale bar represents 200 µm. Scale bar represents 50 µm for the immunofluorescence staining.</p

Oral Administration of ApoB+HSP60 Peptides Induces Tolerance to Both Peptides and attenuates inflammation.

<p>A. Flow cytometry analysis: Percentage of CD25⁺Foxp3⁺ cells (*P = 0.035) and CTLA-4⁺cells (**P<0.0001) within the CD4 population in spleen at the end of the study (n = 6 per group). B. Relative mRNA expression of IFN-γ (*** P<0.001), TGF-β (*** P<0.005), Foxp3 (*** P<0.005), and CTLA-4(*** P<0.005) in the ascending aorta quantified by RT-PCR analysis and normalized to GAPDH. Fold-changes in their expression in ApoB+HSP60-tolerized mice relative to controls are shown (n = 5 per group). C. Splenic effector cells were generated from ApoB/Ldlr^−/− mice immunized subcutaneously with the peptides. Addition of purified Treg cells from oral tolerant mice is indicated at different ratios to effector cells. Proliferation of effector-cell alone is indicated by the white bar; proliferation index represents the percentage carboxyfluorescein succinimidyl ester reduction in culture stimulated with ApoB or HSP60 peptides (10 µg/mL) relative to unstimulated culture (n = 4 per group). *P<0.05 D. Representative photomicrographs showing immunofluorescence staining of aortic sinus sections with CD68 (red) and its quantitative analysis (n = 10 per group). *P = 0.02. E. Representative photomicrographs showing immunofluorescence staining of aortic sinus sections with TNF-α (red) and its quantitative analysis (n = 6 per group). *P = 0.04. F. Representative photomicrographs showing immunofluorescence staining of aortic sinus sections with TGF-β (red) and its quantitative analysis (n = 6 per group). *P = 0.03. Scale bar represents 50 µm for the immunofluorescence staining.</p